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Il 22 1h8pwsr

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IL-22 (1H8PWSR) is a lab equipment product offered by Thermo Fisher Scientific. It is a recombinant protein that serves as a research tool. The core function of this product is to facilitate research and analysis in relevant scientific fields. No further details can be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

Fixation and permeabilization buffer, by BioLegend (2 mentions) Epcam g8, by BioLegend (2 mentions) Fc block, by Thermo Fisher Scientific (2 mentions) Il 13 ebio13a, by Thermo Fisher Scientific (2 mentions) Cell stimulation cocktail, by Thermo Fisher Scientific (2 mentions) Fixable live dead stain, by Thermo Fisher Scientific (2 mentions) Cd45 30 f11, by BioLegend (2 mentions) Dithiothreitol (dtt), by Merck Group (2 mentions) Dnase, by Merck Group (2 mentions) Dispase, by Merck Group (2 mentions) Facsaria 2, by BD (2 mentions) Facscanto 2, by Tree Star (2 mentions) Facsaria 3, by Tree Star (2 mentions) Thy1.2 53 2 1, by Thermo Fisher Scientific (2 mentions) Isotype matched antibodies, by Thermo Fisher Scientific (2 mentions) Ifn γ xmg1.2, by Thermo Fisher Scientific (2 mentions) Il17a 17b7, by Thermo Fisher Scientific (2 mentions) Ter119, by BioLegend (1 mentions) Cd11b m1 70, by Thermo Fisher Scientific (1 mentions) Rb6 8c5, by BioLegend (1 mentions)

8 protocols using il 22 1h8pwsr

1

Isolation and Characterization of Colonic Immune Cells

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Lamina propria cells from colonic tissue were harvested as before8 (link). For intracellular cytokine staining, cells were stimulated using the eBioscience cell stimulation cocktail for 4 h at 37 °C. Cells were fixed and permeabilized using the Biolegend fixation and permeabilization buffer. The following antibodies (clones) were used for staining: CD45 (30-F11), CD19 (6D5), CD11b (M1/70), CD90.2 (53–2.1), CD3 (145–2C11), TCR-β (H57–597), CD4 (RM4–5), CD8 (53–6.7), Gr-1 (RB6–8C5), MHCII (M5/114.15.2), CD11c (N418), CD103 (2E7), IFN-γ (XMG1.2) from Biolegend and IL-22 (1H8PWSR), and IL-13 (eBio13A) from eBioscience. All samples were blocked with Fc block (TruStainfcx) and stained with a fixable live dead stain (Invitrogen). FlowJo v.10 was used to analyze flow cytometry data. Intestinal epithelial cells (IECs) were harvested by incubation at 37°C with 2mM DTT (Sigma) followed by two incubations with 5mM EDTA and then digested with Dispase (Sigma) and DNase (Sigma). IECs were sorted as PI−CD45−EpCam+ using the following antibody clones: CD45 (30-F11) and EpCam (G8.8) from Biolegend on an FACSAria II (BD Biosciences). Purity of IECs was ≥ 95%.
Neil J.A., Matsuzawa-Ishimoto Y., Kernbauer-Hölzl E., Schuster S.L., Sota S., Venzon M., Dallari S., Galvao Neto A., Hine A., Hudesman D., Loke P., Nice T.J, & Cadwell K. (2019). IFN-I and IL-22 mediate protective effects of intestinal viral infection. Nature microbiology, 4(10), 1737-1749.
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2

Comprehensive Flow Cytometry Analysis

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Flow cytometry was performed using a FACSCanto II or FACSAria III and analyzed using FlowJo software (TreeStar). Dead cells were excluded with 7-AAD staining. Non-specific Ab binding was blocked with anti-CD16/32 Ab. Fluorescence-conjugated mAb against CD11b (M1/70), CD11c (N418), Gr-1 (RB6-8C5), F4/80 (BM8), Ly6C (AL-21), MHC class II I-Ab (AF6-120.1), CD103 (2E7), CD45 (30-F11), CD3 (145-2C11), CD4 (GK1.5), NKp46 (29A1.4), Thy-1.2 (53-2.1), and IL-22 (1H8PWSR) were from eBioscience. Isotype-matched antibodies (eBioscience) were used for control staining. All antibodies are used in 1:200 dilution in 1 × 106 cells/100 μl except Gr-1, Ly6C, I-Ab, and CD45 (used in 1:500 dilution).
Seo S.U., Kuffa P., Kitamoto S., Nagao-Kitamoto H., Rousseau J., Kim Y.G., Núñez G, & Kamada N. (2015). Intestinal macrophages arising from CCR2+ monocytes control pathogen infection by activating innate lymphoid cells. Nature Communications, 6, 8010.
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3

Isolation and Analysis of Innate Lymphoid Cells

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The SI of P1 mice were microdissected, minced, and digested with 2 mg ml−1 collagenase D (Roche). Cell suspensions were passed through a 70-μm cell strainer and mononuclear cells were isolated. Cells were pre-incubated with anti-mouse CD16/CD32 for blockade of Fc receptors, and incubated with the appropriate monoclonal Ab conjugates. Propidium iodide (Sigma-Aldrich) or DAPI (4′,6-diamidino-2-phenylindole) (Invitrogen) was used to distinguish live cells from dead cells during cell analysis. Stained cells were analyzed on a FACS Canto or LSRII machine using the Diva software (BD Bioscience). Data were analyzed with FlowJo software (TreeStar). ILC3 cells were defined as CD45+Lin−Thy1+Sca-1hi as described11 (link). The following fluorochrome-conjugated anti-mouse antibodies were used at indicated dilutions: Thy-1.2 (53-2.1, 1:200), Sca-1 (D7, 1:200), CD45 (30-F11, 1:200), CD11b (M1/70, 1:200), IL-22 (1H8PWSR, 1:100), IL-17A (eBio17B7, 1:100) were from eBioscience; lineage cocktail (CD3, B220, CD11b, Gr-1, Ter119−) (145-2C11, RA3-6B2, M1/70, RB6-8C5, TER-119, 1:100) and CX3CR1 (SA011F11, 1:200) were from BioLegend.
Chen L., Strohmeier V., He Z., Deshpande M., Catalan-Dibene J., Durum S.K., Moran T.M., Kraus T., Xiong H., Faith J.J., Sodhi C.P., Hackam D.J., Lira S.A, & Furtado G.C. (2019). Interleukin 22 disrupts pancreatic function in newborn mice expressing IL-23. Nature Communications, 10, 4517.
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4

Immune Cell Phenotyping Protocol

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Antibodies against mouse CD3 (145-2C11), CD4 (GK1.5), CD45 (30-F11), Foxp3 (FJK-16s), IL-10 (JES5-16E3), CD11c (N418), CD11b (M1/70), I-Ab (25-9-17), CD90.1 (HIS51), V alpha 2 TCR (B20.1), V beta 5.1/5.2 TCR (MR9-4), IFN-γ (XMG1.2), IL-22 (1H8PWSR) and IL17A (17B7) were purchased from eBioscience and Biolegend. GPR81 and actin antibodies were purchased from Sigma.
Ranganathan P., Shanmugam A., Swafford D., Suryawanshi A., Bhattacharjee P., Hussein M.S., Koni P.A., Prasad P.D., Kurago Z.B., Thangaraju M., Ganapathy V, & Manicassamy S. (2018). GPR81, a cell-surface receptor for lactate, regulates intestinal homeostasis and protects mice from experimental colitis. Journal of immunology (Baltimore, Md. : 1950), 200(5), 1781-1789.
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5

Multicolor Flow Cytometric Analysis

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Flow cytometry was performed using a FACSCanto II or FACSAria III and analysed using FlowJo software (TreeStar). Dead cells were excluded with 7-AAD staining. Non-specific antibody binding was blocked with anti-CD16/32 antibody. Fluorescence-conjugated mAb against CD11b (M1/70), CD11c (N418), Gr-1 (RB6-8C5), F4/80 (BM8), Ly6C (AL-21), MHC class II I-Ab (AF6-120.1), CD103 (2E7), CD45 (30-F11), CD3 (145-2C11), CD4 (GK1.5), NKp46 (29A1.4), Thy-1.2 (53-2.1) and IL-22 (1H8PWSR) were from eBioscience. Isotype-matched antibodies (eBioscience) were used for control staining. All antibodies are used in 1:200 dilution in 1 × 106 cells per 100 μl except Gr-1, Ly6C, I-Ab and CD45 (used in 1:500 dilution).
Seo S.U., Kuffa P., Kitamoto S., Nagao-Kitamoto H., Rousseau J., Kim Y.G., Núñez G, & Kamada N. (2015). Intestinal macrophages arising from CCR2+ monocytes control pathogen infection by activating innate lymphoid cells. Nature Communications, 6, 8010.
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6

Isolation and Characterization of Colonic Immune Cells

Cited 1 time
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Lamina propria cells from colonic tissue were harvested as before8 (link). For intracellular cytokine staining, cells were stimulated using the eBioscience cell stimulation cocktail for 4 h at 37 °C. Cells were fixed and permeabilized using the Biolegend fixation and permeabilization buffer. The following antibodies (clones) were used for staining: CD45 (30-F11), CD19 (6D5), CD11b (M1/70), CD90.2 (53–2.1), CD3 (145–2C11), TCR-β (H57–597), CD4 (RM4–5), CD8 (53–6.7), Gr-1 (RB6–8C5), MHCII (M5/114.15.2), CD11c (N418), CD103 (2E7), IFN-γ (XMG1.2) from Biolegend and IL-22 (1H8PWSR), and IL-13 (eBio13A) from eBioscience. All samples were blocked with Fc block (TruStainfcx) and stained with a fixable live dead stain (Invitrogen). FlowJo v.10 was used to analyze flow cytometry data. Intestinal epithelial cells (IECs) were harvested by incubation at 37°C with 2mM DTT (Sigma) followed by two incubations with 5mM EDTA and then digested with Dispase (Sigma) and DNase (Sigma). IECs were sorted as PI−CD45−EpCam+ using the following antibody clones: CD45 (30-F11) and EpCam (G8.8) from Biolegend on an FACSAria II (BD Biosciences). Purity of IECs was ≥ 95%.
Neil J.A., Matsuzawa-Ishimoto Y., Kernbauer-Hölzl E., Schuster S.L., Sota S., Venzon M., Dallari S., Galvao Neto A., Hine A., Hudesman D., Loke P., Nice T.J, & Cadwell K. (2019). IFN-I and IL-22 mediate protective effects of intestinal viral infection. Nature microbiology, 4(10), 1737-1749.
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7

Characterizing Skin Cell Immune Phenotypes

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Skin cells were preincubated with FcγR-specific blocking mAb (2.4G2) and washed before staining with the following monoclonal antibodies (mAbs): CD3 (17A2), CD4 (GK1.5), CD45 (30F11) and δ TCR (ebioGL3) from eBioscience, CD11b (M1/70), and CD117 (2B8) from Biolegend and anti-IgE (R35–72) from BD Biosciences. For cytokine staining, cells were stimulated with Ionomycin (0.5ug/ml; Sigma), Phorbol 12,13-dibutyrate (1ug/ml; Sigma), Brefeldin A (eBioscience), Monensin (eBioscience) in complete RPMI for 3 hours before surface staining. Then, cells were fixed and permeabilized (BD Biosciences Cytofix/Cytoperm) and stained in permeabilization solution with IL-22 (1H8PWSR), from Ebiosciences. Cells were analyzed by flow cytometry using an LSRFortessa machine (BD Biosciences). The data was analyzed with FlowJo software.
Leyva-Castillo J.M., Sun L., Wu S.Y., Rockowitz S., Sliz P, & Geha R. (2022). Single cell transcriptome profile of mouse skin undergoing antigen driven allergic inflammation recapitulates findings in atopic dermatitis skin lesions. The Journal of allergy and clinical immunology, 150(2), 373-384.
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8

Murine Immune Cell Characterization

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Antibodies against mouse CD3 (145-2C11), CD4 (GK1.5), CD8a (53-6.7), NK1.1 (PK136), CD335 (29A1.4), CD19 (1D3), CD45 (30-F11), Foxp3 (FJK-16s), IL-10 (JES5-16E3), CD11c (N418), CD11b (M1/70), I-Ab ( 25-9-17), CD90.1 (HIS51), V alpha 2 TCR (B20.1), V beta 5.1/5.2 TCR (MR9-4), IFN-γ (XMG1.2), IL-22 (1H8PWSR) and IL17A (17B7) were purchased from eBioscience. PPARα antibody was obtained from Cell Signaling Technology. PPARα antagonist GW6471 and orally active PPARα agonists GW7647 were purchased from Tacoris. OVA323−339 (ISQVHAAHAEINEAGR) peptide was purchased from Anaspec.
Manoharan I., Suryawanshi A., Hong Y., Ranganathan P., Shanmugam A., Ahmad S., Swafford D., Manicassamy B., Ramesh G., Koni P.A., Thangaraju M, & Manicassamy S. (2016). Homeostatic PPARα signaling limits inflammatory responses to commensal microbiota in the intestine. Journal of immunology (Baltimore, Md. : 1950), 196(11), 4739-4749.
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